LOCUS V044455 9191 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044455 VERSION V044455 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3552 /codon_start=1 /gene="" /note="" /product="human lysosome-associated membrane glycoprotein 1" /protein_id="" /transl_table=1 /translation="MAAPGSARRPLLLLLLLLLLGLMHCASA,AMFMVKNGNGTACIMA NFSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLN FTRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVH MNNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDK YNVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHS EGTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCN AEEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLI VLIAYLVGRKRSHAGYQTI" /label="LAMP1" CDS 3595..4305 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /transl_table=1 /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" /label="mTagBFP2" promoter 4356..4567 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4580..4848 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4881..5279 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 5286..5874 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5948..6181 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6253..6374 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6393..6528 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6566..6582) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6590..6606 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6614..6644) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6659..6680 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6968..7556) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7727..8587) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8588..8692) /gene="bla" /label="AmpR promoter" primer_bind 9166..9182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggcggcc cccggcagcg cccggcgacc cctgctgctg 2341 ctactgctgt tgctgctgct cggcctcatg cattgtgcgt cagcagcaat gtttatggtg 2401 aaaaatggca acgggaccgc gtgcataatg gccaacttct ctgctgcctt ctcagtgaac 2461 tacgacacca agagtggccc taagaacatg acctttgacc tgccatcaga tgccacagtg 2521 gtgctcaacc gcagctcctg tggaaaagag aacacttctg accccagtct cgtgattgct 2581 tttggaagag gacatacact cactctcaat ttcacgagaa atgcaacacg ttacagcgtc 2641 cagctcatga gttttgttta taacttgtca gacacacacc ttttccccaa tgcgagctcc 2701 aaagaaatca agactgtgga atctataact gacatcaggg cagatataga taaaaaatac 2761 agatgtgtta gtggcaccca ggtccacatg aacaacgtga ccgtaacgct ccatgatgcc 2821 accatccagg cgtacctttc caacagcagc ttcagccggg gagagacacg ctgtgaacaa 2881 gacaggcctt ccccaaccac agcgccccct gcgccaccca gcccctcgcc ctcacccgtg 2941 cccaagagcc cctctgtgga caagtacaac gtgagcggca ccaacgggac ctgcctgctg 3001 gccagcatgg ggctgcagct gaacctcacc tatgagagga aggacaacac gacggtgaca 3061 aggcttctca acatcaaccc caacaagacc tcggccagcg ggagctgcgg cgcccacctg 3121 gtgactctgg agctgcacag cgagggcacc accgtcctgc tcttccagtt cgggatgaat 3181 gcaagttcta gccggttttt cctacaagga atccagttga atacaattct tcctgacgcc 3241 agagaccctg cctttaaagc tgccaacggc tccctgcgag cgctgcaggc cacagtcggc 3301 aattcctaca agtgcaacgc ggaggagcac gtccgtgtca cgaaggcgtt ttcagtcaat 3361 atattcaaag tgtgggtcca ggctttcaag gtggaaggtg gccagtttgg ctctgtggag 3421 gagtgtctgc tggacgagaa cagcatgctg atccccatcg ctgtgggtgg tgccctggcg 3481 gggctggtcc tcatcgtcct catcgcctac ctcgtcggca ggaagaggag tcacgcaggc 3541 taccagacta tcggatccgg cggtggagga agcggcggag gtggatctct cgagatggtg 3601 tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat ggagggcacc 3661 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc 3721 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg 3781 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac 3841 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac 3901 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac 3961 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc 4021 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac 4081 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat 4141 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga 4201 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg 4261 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattaagg atccgcggcc 4321 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt 4381 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg 4441 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga 4501 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag 4561 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg 4621 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg 4681 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc 4741 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc 4801 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc 4861 ggcgcctacg ctaggccacc atggccaagc ctttgtctca agaagaatcc accctcattg 4921 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg 4981 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg 5041 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga 5101 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc 5161 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac 5221 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag 5281 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 5581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 5641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 5701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 5761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 5821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta 5881 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 5941 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact 6001 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 6061 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 6121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 6181 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca 6241 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 6301 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 6361 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg 6421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 6481 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca 6541 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 6601 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 6661 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 6721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 6781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 6841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 6901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 6961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 7021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 7081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 7141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 7201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 7261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 7321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 7381 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 7441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 7501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 7561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 7621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 7681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 7741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 7801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 7861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 7921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 7981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 8041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 8101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 8161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 8221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 8281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 8341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 8401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 8461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 8521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 8581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 8641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 8701 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 8761 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 8821 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 8881 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 8941 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg 9001 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag 9061 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 9121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 9181 gtgccaagct g //